This study was aimed to investigate the effects of oxaliplatin on human multiple myeloma cell line RPMI-8226 and its mechanism. The proliferation inhibitory rate of RPMI8226 cells was assayed by MTT, the morphological changes of RPMI-8226 cells were observed by inverted fluorescent microscopy and transmission electron microscopy, the apoptosis rate and the cell cycle distribution of RPMI-8226 cells were detected by flow cytometry, The effects of oxaliplatin on the expression of Bcl-2, caspase-8, caspase-3 mRNA were tested by RT-PCR, Bcl-2 protein expression of RPMI-8226 cells was analyzed by Western blot. The results showed that oxaliplatin could inhibit the proliferation of RPMI-8226 cells in time and dose-dependent manners. Cell number in oxaliplatin group was significantly less than that in control group under light microscope, and the growth arrangement was irregular, apoptotic cells could be seen. Under electron microscope, typical apoptotic morphological and ultrastructural changes could be observed. Flow cytometry results showed that oxaliplatin could induce apoptosis of RPMI-8226 cells, the difference have statistical significance (P < 0.05). Oxaliplatin mainly arrested RPMI-8226 cells in the S phase (P < 0.05). The expression of Bcl-2 mRNA did not have apparent change, while the expression of caspase-8, caspase-3 mRNA increased (P < 0.05). Western blot results suggested that the expression of Bcl-2 protein had no obvious change. It is concluded that the oxaliplatin can induce apoptosis of RPMI-8226 cells, activating the death receptor pathway and arresting cell cycle may be two of the related mechanisms, Bcl-2 gene has unobservable effects in the process of oxaliplatin-induced cell apoptosis.
Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Multiple Myeloma ; metabolism ; pathology ; Organoplatinum Compounds ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism

This study was aimed to investigate the effects of docetaxel on proliferation and apoptosis of multiple myeloma cell line RPMI8226 and its mechanism. The inhibitory rate of multiple myeloma cells was detected by MTT, the morphological and ultrastructural changes of RPMI8226 cells were observed by using inverted fluorescent microscope and transmission electron microscope, the apoptosis-inducing effect of docetaxel on RPMI-8226 cells was determined by flow cytometry with Annexin-V FITC/PI staining, the cell distribution in cell cycle of RPMI-8226 cells was assayed using flow cytometry with PI staining; the effect of docetaxel on expression of BCL-2, caspase-8, caspase-3 mRNA was detected by semiquantitative RT-PCR, the expression changes of BCL-2 protein in RPMI-8226 cells before and after treatment with docetaxel were measured by using Western blot. The results indicated that 0.25 - 8.0 µg/ml docetaxel obviously inhibited the proliferation of RPMI-8226 cells in both time- and dose-dependent manners. Cell number of docetaxel-treated group was significantly less than control group under inverted fluorescent microscope, and the cell arrangement was irregular, necrotic cells could be seen occasionally. By transmission electron microscope, the morphological and ultrastructural changes of cell typical apoptosis could be observed, a few necrotic cells could be captured, too. Compared with control group, docetaxel induced the apoptosis of RPMI-8226 cell line (P < 0.01). Docetaxel mainly arrested RPMI-8226 cells in the G(2)/M phase (P < 0.01). The expression of BCL-2 mRNA decreased (P < 0.05), while the mRNA expression of caspase-8 and caspase-3 increased (P < 0.05). Western blot indicated that BCL-2 protein expression also decreased (P < 0.05). It is concluded that docetaxel can inhibit the proliferation of RPMI-8226 cells by inducing cell apoptosis. Activation of the mitochondrial and death receptor pathways of apoptosis may be involved in the docetaxel-induced apoptosis, cell cycle arrest may also play an important role in the apoptosis mechanism.
Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Multiple Myeloma ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Taxoids ; pharmacology

OBJECTIVETo explore the prevalence of IDH gene (IDH1 and IDH2) mutations, types of mutations in patients with acute myeloid leukemia (AML), correlation with the internal tandem duplication(ITD) mutation of FLT3 gene, NPM1 gene mutation and some clinical characteristics.

METHODSThe mutations of IDH1 and IDH2 gene at exon 4, NPM1 gene at exon 12 and FLT3-ITD at exon 14 and 15 in 163 newly diagnosed AML patients were detected by PCR amplification followed by direct sequencing of genomic DNA.

RESULTS(1) IDH mutations were found in 25 patients (25/163), and all were heterozygous, of which IDH1 in 7 patients (4.29%) and IDH2 in 18 (11.04%). A total of 4 types of IDH1 mutations were identified (c.395G→A, p.R132H, n = 4; c.394C→A, p.R132S, n = 1; c.394C→G, p.R132G, n = 1; c.315C→T, n = 1). The IDH1 mutation caused substitutions of residue R132 except for one (c.315C→T). All IDH2 mutations caused changes of R140 (c.419G→A, p.R140Q, n = 18). The incidence of IDH2 mutation was significantly higher than that of IDH1 mutation (11.0% v 4.3%, P = 0.022). Both IDH1 and IDH2 mutation were detected in one patient, while IDH1 was synonymous substitution (c.315C→T). IDH-mutated cases showed a significantly higher frequency of concurrent FLT3-ITD mutation compared with wildtype cases (34.6% vs 11.9%, P = 0.003), so did IDH mutations concurrent NPM1 mutation vs NPM1 wildtype (28.1% vs 12.7%, P = 0.033), of which the frequency of concurrent NPM1 and FLT-ITD mutations cases with the IDH mutation was significantly higher than that of NPM1 and FLT-ITD negative (45.5% vs 11.7%, P = 0.002). IDH mutation incidence was significantly higher in normal karyotype cases than in abnormal ones (20.5% vs 5.8%, P = 0.020). Patients with IDH mutations were significantly older than wildtype patients(P < 0.001), whereas, there were no statistically significant differences in gender, peripheral blood (PB) count at diagnosis between two groups.

CONCLUSIONSThe incidence of IDH mutation is higher in patients with de novo AMLs, of which IDH2 mutation more frequently, and the patients associated with older age, normal karyotype at diagnosis. IDH mutation has a strong association with NPM1 and FLT3-ITD mutations, suggesting that IDH mutation has synergistic effect with the latter gene on leukemogenesis.

Adolescent ; Adult ; Aged ; DNA Mutational Analysis ; Female ; Genotype ; Humans ; Isocitrate Dehydrogenase ; genetics ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Young Adult

OBJECTIVETo understand the situation of institutional delivery of rural pregnant women in Guangxi Autonomous Region in the period of 1998 - 2003 and to identify the determinants on institutional delivery utilization.

METHODSUsing Andersen's behavioral model as analytical framework and Guangxi databank of the 3rd National Health Service Survey as data source, we described the status of institutional delivery with the rural women having had live birth history in the period of 1998 - 2003 as subjects, while and the univariate analysis and multivariate logistic analysis were done to identify determinants of institutional delivery utilization.

RESULTSAmong a total number of 407 women with live birth history, 39.80 percent of them delivered at the health-care facilities. The rate of institutional delivery increased annually in 1998 - 2003 (P< 0.0001). The proportion of delivery in township health centers increased and the proportion of home delivery decreased by year (P< 0.0001). Results from both univariate and multivariate analysis showed that parity, education background of women, type of drinking water, time needed to get to the nearest healthcare facilities by the most convenient traffic,frequency of prenatal checkup, together with whether or not being advocated on institutional delivery etc. were determinants of delivery utilization. The OR value were 1.749 for multipara, 1.995 for those going to the nearest healthcare facilities by the most convenient traffic in less than 10 minutes, 3.011 for those drinking tap water, 5.435 for those with the education of high school, 29.149 for those with over 5 times in terms of frequency of prenatal checkup and 37.822 for those being advocated on institutional delivery.

CONCLUSIONSocio-economic situation, status of maternal health care and parity made main contribution to institutional delivery and skilled birth attendance for women in rural Guangxi.

China ; Choice Behavior ; Delivery, Obstetric ; methods ; utilization ; Female ; Hospitalization ; statistics & numerical data ; Humans ; Models, Statistical ; Pregnancy ; Rural Population

OBJECTIVETo investigate the composition and morphology of the stones in the enlarged prostatic utricle (EPU).

METHODSWe took out 36 EPU stones from 11 patients by transurethral fenestration between 1992 and 2011, and analyzed the stones by scanning electron microscopy, x-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIS).

RESULTSUnder the scanning electron microscope, all the EPU stones were constituted of many intensive minicrystals and amorphous matrix. XRD and FTIS revealed that all were hydroxyapatite crystal.

CONCLUSIONEPU stones belong to the category of prostatic pseudo-calculi, whose formation is ascribed not to the abnormal change of urine composition, but to the continuous secretion, absorption and concentration of EPU liquid and ablated epithelial cells from the EPU.

Calculi ; chemistry ; Durapatite ; chemistry ; Humans ; Male ; Prostate ; chemistry ; pathology ; Prostatic Diseases ; pathology ; physiopathology

OBJECTIVETo analyze the correlation between clinical features and cytogenetic finding of 45 adult patients with acute lymphoblastic leukemia (ALL), and to assess the value of chromosomal examination for the diagnosis and prognosis.

METHODSFluorescence in situ hybridization (FISH) was utilized for detecting the BCR/ABL fusion gene and P53 gene. Median survival time (MST) of patients was compared using Log-rank test.

RESULTSRespectively, the MST of patients with white blood cell count (WBC) ≤30 × 10(9)/L, normal karyotype, or without a Philadelphia chromosome were significantly greater than those with WBC > 30 × 10(9)/L, abnormal karyotype or Philadelphia chromosome (P< 0.05).

CONCLUSIONWBC, karyotype abnormalities and presence of Philadelphia chromosome are independent factors for the prognosis of ALL in adult patients.

Abnormal Karyotype ; Adult ; Aged ; Cytogenetic Analysis ; methods ; Female ; Fusion Proteins, bcr-abl ; genetics ; Genes, p53 ; Humans ; Male ; Middle Aged ; Philadelphia Chromosome ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; genetics

OBJECTIVETo investigate the prognostic value of t(11; 18) (q21; q21) in gastric mucosa-associated lymphoid tissue lymphoma.

METHODSA cohort of thirty-six gastric mucosa-associated lymphoid tissue lymphoma patients who were pathologically identify diagnosis from January 1994 to June 2004 were followed up retrospectively and studied using fluorescence in situ hybridization(FISH) technique to detect t(11; 18) (q21; q21) chromosomal translocation on preservative paraffin specimen.

RESULTSAmong thirty-six patients, fifteen (41.67%) were positive for t (11; 18) (q21; q21). All but one were followed up to March 2010, general median survival time (MST) was 87 months. The MST were 43 and 130 months for t(11; 18) positive and negative patients, respectively. The MST between these two groups was notably different (chi-square=29.57, P< 0.01).

CONCLUSIONt(11; 18) (q21; q21) is important prognostic factor for gastric mucosa-associated lymphoid tissue lymphoma.

Adult ; Aged ; Aged, 80 and over ; Chromosomes, Human, Pair 11 ; Chromosomes, Human, Pair 18 ; Cohort Studies ; Female ; Follow-Up Studies ; Gastric Mucosa ; pathology ; Humans ; Lymphoma, B-Cell, Marginal Zone ; genetics ; pathology ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Translocation, Genetic

This study was purposed to explore the mechanisms of cladribine (2-CdA)-inducing apoptosis of multiple mycloma RPMI 8226 cells. The MTT method was used to determine cell proliferation after being treated with 2-CdA. Apoptosis and cell cycle progression were examined by flow cytometry. Transmission electron microscopy was used to observe ultrastructural changes of RPMI 8226 cells. RT-PCR and Western blot were used to analyze the mRNA and protein expression levels of BCL-2, MCL-2 and caspase-3 respectively. The results showed that the 2-CdA inhibited proliferation of RPMI 8226 cells in time and dose-dependent manner. Typical apoptotic morphological and ultrastructure changes could be observed by electron microscopy. Flow cytometry showed that 2-CdA induced myeloma cell apoptosis and arrested myeloma cells in the G2/M phase. The mRNA expression of BCL-2 and MCL-1 decreased but that of caspase-3 not apparently changed. Western blot results suggested that the change trend of BCL-2 MCL-1 and caspase-3 was the same as result of RT-PCR. It is concluded that 2-CdA exhibits inhibitory effects on RPMI 8226 cells in vitro. Activating the mitochondrial and death receptor pathways of apoptosia may be the potential mechanism, meanwhile, the cell cycle arrest may also play a critical role in apoptosis.
Apoptosis ; drug effects ; Caspase 3 ; Cell Division ; Cell Line, Tumor ; Cell Proliferation ; Cladribine ; pharmacology ; Humans ; Multiple Myeloma ; pathology ; Proto-Oncogene Proteins c-bcl-2

This study was aimed to investigate the effects of mitoxantrone on proliferation and apoptosis of human multiple myeloma cell line RPMI-8226 and its mechanism. The inhibitory rate of RPMI8226 cells proliferation was assayed by MTT, the morphological changes of RPMI-8226 cells were observed by inverted flurescent microscopy and transmission electron microscopy, the apoptosis rate and the cell cycle distribution of RPMI-8226 cells were detected by flow cytometry. The effects of mitoxantrone on the expression of BCL-2, BAX, caspase-3 mRNA were detected by RT-PCR, the BCL-2, BAX, caspase-3 protein expression of RPMI-8226 cells was analyzed by Western blot. The results showed that mitoxantrone could inhibit the proliferation of RPMI-8226 cells in time- and dose-dependent manners. Light microscopy showed that the cell number in mitoxantrone group was significantly less than that in control group and the cell growth arrangement was irregular, apoptotic cells could be seen. Under electron microscope, typical apoptotic morphological and ultrastructural changes could be observed, these results confirmed that the mitoxantrone could induce apoptosis of RPMI-8226 cells, the difference have statistical significance (P < 0.05). The 1.0 µg/ml low concentration of mitoxantrone mainly arrested RPMI-8226 cells in the G2/M phase(P < 0.05), and the 2.0 µg/ml high concentration of mitoxantrone mainly arrested RPMI-8226 cells in the S phase (P < 0.05). The expression of BCL-2 mRNA decreased (P < 0.05),while the expression of BAX, caspase-3 mRNA increased (P < 0.05). Western blot indicated that BCL-2 protein expression also decreased (P < 0.05) and BAX, caspase-3 protein expression increased. It is concluded that the mitoxantrone can inhibit the proliferation of RPMI-8226 cells by inducing cell apoptosis. Activation of the mitochondrial and death receptor pathways of apoptosis may be involved in the mitoxantrone-induced apoptosis, the cell cycle arrest may also play an important role in the apoptosis mechanism.
Apoptosis ; drug effects ; Caspase 3 ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Mitoxantrone ; pharmacology ; Multiple Myeloma ; pathology ; Proto-Oncogene Proteins c-bcl-2

This study was aimed to investigate the effects of sorafenib on proliferation and apoptosis of MM cell line RPMI-8226, and to explore the its potential anti-tumor mechanism. The inhibitory rate of multiple myeloma cell proliferation was tested by MTT. Transmission electron microscopy was used to observe morphological and ultrastructural changes of RPMI-8226 cells treated with sorafenib. The effects of sorafenib on the apoptosis and cell cycle of RPMI-8226 cells was detected by flow cytometry. The effects of sorafenib on the expression of caspase-3, BCL-2 and MCL-1 mRNA and protein were assayed by RT-PCR and Western blot respectively. The results showed that sorafenib (0-10.0 µmol/L) could obviously inhibit the proliferation of RPMI-8226 cells in time and dose-dependent manner. Flow cytometry results showed that sorafenib could induce apoptosis of RPMI-8226 cells, the difference was statistical significance (P < 0.05). Sorafenib mainly arrested RPMI-8226 cells in the G1 phase (P < 0.05). Typical apoptotic morphological and ultrastructural changes of MM cells could be observed under transmission electron microscope, Examination of cellular signaling pathways showed that sorafenib induced upregulation of cleaved-caspase-3 expression, and simultaneous downregulation of BCL-2 and MCL-1 expression. It is concluded that sorafenib displays anti-myeloma activity. Activating the death receptor pathway and arresting cell cycle may be two of the relatated mechanisms.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Multiple Myeloma ; drug therapy ; pathology ; Niacinamide ; analogs & derivatives ; pharmacology ; Phenylurea Compounds ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; Signal Transduction